Azaindoles and methods of use thereof

ABSTRACT

Disclosed are compounds according to Formula (I) or (II), and pharmaceutical compositions comprising them. Also disclosed are therapeutic methods, e.g., of treating kidney diseases, using the compounds of Formula (I) or (II).

RELATED APPLICATION

This application claims benefit of priority to U.S. Provisional PatentApplication Ser. No. 62/541,461, filed Aug. 4, 2017.

BACKGROUND

Proteinuria is a condition in which an excessive amount of protein inthe blood leaks into the urine. Proteinuria can progress from a loss of30 mg of protein in the urine over a 24-hour period (calledmicroalbuminuria) to >300 mg/day (called macroalbuminuria), beforereaching levels of 3.5 grams of protein or more over a 24-hour period,or 25 times the normal amount. Proteinuria occurs when there is amalfunction in the kidney's glomeruli, causing fluid to accumulate inthe body (edema). Prolonged protein leakage has been shown to result inkidney failure. Nephrotic Syndrome (NS) disease accounts forapproximately 12% of prevalent end stage renal disease cases at anannual cost in the United States of more than $3 billion. Approximately5 out of every 100,000 children are diagnosed with NS every year and 15out of every 100,000 children are living with it today. For patients whorespond positively to treatment, the relapse frequency is extremelyhigh. 90% of children with Nephrotic Syndrome will respond to treatment,however, an estimated 75% will relapse. Therefore, more effectivemethods of treating, or reducing risk of developing, kidney disease,e.g., proteinuria, are desirable.

Mammalian TRP channel proteins form six-transmembrane cation-permeablechannels that may be grouped into six subfamilies on the basis of aminoacid sequence homology (TRPC, TRPV, TRPM, TRPA, TRPP, and TRPML). Recentstudies of TRP channels indicate that they are involved in numerousfundamental cell functions and are considered to play an important rolein the pathophysiology of many diseases. Many TRPs are expressed inkidney along different parts of the nephron and growing evidence suggestthat these channels are involved in hereditary, as well as acquiredkidney disorders. TRPC6, TRPM6, and TRPP2 have been implicated inhereditary focal segmental glomerulosclerosis (FSGS), hypomagnesemiawith secondary hypocalcemia (HSH), and polycystic kidney disease (PKD),respectively.

TRPC5 has also been reported to contribute to the mechanisms underlyingregulation of innate fear responses. (J Neurosci. 2014 Mar. 5; 34(10):3653-3667).

Hence, there is a need for additional inhibitors of TRPC5.

SUMMARY

This invention is based, at least in part, on the discovery thatTransient Receptor Potential Cation Channel, subfamily C, member 5(TRPC5) activity abolishes actin stress fibers and diminishes focaladhesion formation, rendering a motile, migratory podocyte phenotype.

In one aspect, the invention relates to methods of treating, or reducingrisk of developing, kidney disease (e.g., proteinuria, microalbuminuria,macroalbuminuria), anxiety, depression, or cancer, in a subject byadministering a therapeutically effective amount of a TRPC5 inhibitor tothe subject.

The methods are effective for a variety of subjects including mammals,e.g., humans and other animals, such as laboratory animals, e.g., mice,rats, rabbits, or monkeys, or domesticated and farm animals, e.g., cats,dogs, goats, sheep, pigs, cows, or horses.

In some embodiments, the methods include administering a small moleculethat inhibits TRPC5.

In some embodiments, the compound of the invention is a compound ofFormula (I) or (II), or a pharmaceutically acceptable salt thereof;

wherein

-   -   R¹ is selected from the group consisting of alkyl; cycloalkyl;        heterocycloalkyl; aryl; heteroaryl; alkylene-aryl;        alkylene-heteroaryl; alkylene-O-aryl; alkylene-N(alkyl)₂;        alkylene-heterocycloalkyl; alkylene-cycloalkyl; —N(alkyl)₂; and        —C(O)-aryl;    -   R² is selected from the group consisting of alkyl; cycloalkyl;        heterocycloalkyl; aryl; heteroaryl; alkylene-N(alkyl)₂;        alkylene-heterocycloalkyl; alkylene-cycloalkyl;        alkylene-heterocycloalkyl; and alkylene-OR′;    -   R³ is independently selected from alkyl, halogen, OMe, OH,        N(Me)₂, CF₃, OCF₃, CHF₂, OCHF₂, and —O-alkylene-OH;    -   R′ is H, methyl, ethyl, or isopropyl; and    -   n is 0, 1, 2, 3, or 4;    -   provided that the compound is not

In one aspect, the invention features a composition, comprising acompound of any one of Formula (I) or (II) or a pharmaceuticallyacceptable salt thereof; and a pharmaceutically acceptable excipient.

In one aspect, the invention features methods of treating, or thereducing risk of developing, a kidney disease, anxiety, or depression,or cancer, comprising administering to a subject in need thereof atherapeutically effective amount of a compound of Formula (I) or (II).

In certain embodiments, a kidney disease is treated or the risk ofdeveloping a kidney disease is reduced. In certain embodiments, a kidneydisease is treated. In certain embodiments, the kidney disease isselected from the group consisting of Focal Segmental Glomerulosclerosis(FSGS), Diabetic nephropathy, Alport syndrome, hypertensive kidneydisease, nephrotic syndrome, steroid-resistant nephrotic syndrome,minimal change disease, membranous nephropathy, idiopathic membranousnephropathy, membranoproliferative glomerulonephritis (MPGN), immunecomplex-mediated MPGN, complement-mediated MPGN, Lupus nephritis,postinfectious glomerulonephritis, thin basement membrane disease,mesangial proliferative glomerulonephritis, amyloidosis (primary), c1qnephropathy, rapidly progressive GN, anti-GBM disease, C3glomerulonephritis, hypertensive nephrosclerosis, and IgA nephropathy.In certain embodiments, the kidney disease is proteinuria. In certainembodiments, the kidney disease is microalbuminuria or macroalbuminuria.

In certain embodiments, the subject is a mammal. In certain embodiments,the mammal is a human.

In some embodiments, the invention comprises administering the compoundof Formula (I) or (II) to a mammal and evaluating an effect of thecompound on calcium transport, wherein a compound that reduces orinhibits calcium transport is a therapeutic agent for treating orreducing risk of developing a kidney disease, anxiety, depression, orcancer.

The invention provides several advantages. The prophylactic andtherapeutic methods described herein are effective in treating kidneydisease, e.g., proteinuria, and have minimal, if any, side effects.Further, methods described herein are effective to identify compoundsthat treat or reduce risk of developing a kidney disease, anxiety,depression, or cancer.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described below. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference in their entirety. In case of conflict, the presentspecification, including definitions, will control. In addition, thematerials, methods, and examples are illustrative only and not intendedto be limiting.

Other features, objects, and advantages of the invention will beapparent from the detailed description, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 tabulates characterization data for representative compounds ofthe invention.

DETAILED DESCRIPTION Definitions

The term “acyl” is art-recognized and refers to a group represented bythe general formula hydrocarbylC(O)—, preferably alkylC(O)—.

The term “acylamino” is art-recognized and refers to an amino groupsubstituted with an acyl group and may be represented, for example, bythe formula hydrocarbylC(O)NH—.

The term “acyloxy” is art-recognized and refers to a group representedby the general formula hydrocarbylC(O)O—, preferably alkyC(O)O—.

The term “alkoxy” refers to an alkyl group, preferably a lower alkylgroup, having an oxygen attached thereto. Representative alkoxy groupsinclude methoxy, trifluoromethoxy, ethoxy, propoxy, tert-butoxy and thelike.

The term “alkoxyalkyl” refers to an alkyl group substituted with analkoxy group and may be represented by the general formulaalkyl-O-alkyl.

The term “alkenyl”, as used herein, refers to an aliphatic groupcontaining at least one double bond and is intended to include both“unsubstituted alkenyls” and “substituted alkenyls”, the latter of whichrefers to alkenyl moieties having substituents replacing a hydrogen onone or more carbons of the alkenyl group. Such substituents may occur onone or more carbons that are included or not included in one or moredouble bonds. Moreover, such substituents include all those contemplatedfor alkyl groups, as discussed below, except where stability isprohibitive. For example, substitution of alkenyl groups by one or morealkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups iscontemplated.

An “alkyl” group or “alkane” is a straight chained or branchednon-aromatic hydrocarbon which is completely saturated. Typically, astraight chained or branched alkyl group has from 1 to about 20 carbonatoms, preferably from 1 to about 10 unless otherwise defined. Examplesof straight chained and branched alkyl groups include methyl, ethyl,n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl,pentyl and octyl. A C₁-C₆ straight chained or branched alkyl group isalso referred to as a “lower alkyl” group.

Moreover, the term “alkyl” (or “lower alkyl”) as used throughout thespecification, examples, and claims is intended to include both“unsubstituted alkyls” and “substituted alkyls”, the latter of whichrefers to alkyl moieties having substituents replacing a hydrogen on oneor more carbons of the hydrocarbon backbone. Such substituents, if nototherwise specified, can include, for example, a halogen (e.g., fluoro),a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl,or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or athioformate), an alkoxy, a phosphoryl, a phosphate, a phosphonate, aphosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro,an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, asulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or anaromatic or heteroaromatic moiety. In preferred embodiments, thesubstituents on substituted alkyls are selected from C₁₋₆ alkyl, C₃₋₆cycloalkyl, halogen, carbonyl, cyano, or hydroxyl. In more preferredembodiments, the substituents on substituted alkyls are selected fromfluoro, carbonyl, cyano, or hydroxyl. It will be understood by thoseskilled in the art that the moieties substituted on the hydrocarbonchain can themselves be substituted, if appropriate. For instance, thesubstituents of a substituted alkyl may include substituted andunsubstituted forms of amino, azido, imino, amido, phosphoryl (includingphosphonate and phosphinate), sulfonyl (including sulfate, sulfonamido,sulfamoyl and sulfonate), and silyl groups, as well as ethers,alkylthios, carbonyls (including ketones, aldehydes, carboxylates, andesters), —CF₃, —CN and the like. Exemplary substituted alkyls aredescribed below. Cycloalkyls can be further substituted with alkyls,alkenyls, alkoxys, alkylthios, aminoalkyls, carbonyl-substituted alkyls,—CF₃, —CN, and the like.

Unless otherwise specified, “alkylene” by itself or as part of anothersubstituent refers to a saturated straight-chain or branched divalentgroup having the stated number of carbon atoms and derived from theremoval of two hydrogen atoms from the corresponding alkane. Examples ofstraight chained and branched alkylene groups include —CH₂— (methylene),—CH₂—CH₂-(ethylene), —CH₂—CH₂—CH₂— (propylene), —C(CH₃)₂—,—CH₂—CH(CH₃)—, —CH₂—CH₂—CH₂—CH₂—CH₂—CH₂—CH₂—CH₂—CH₂— (pentylene),—CH₂—CH(CH₃)—CH₂—, and —CH₂—C(CH₃)₂—CH₂—.

The term “C_(x-y)” when used in conjunction with a chemical moiety, suchas, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant toinclude groups that contain from x to y carbons in the chain. Forexample, the term “C_(x-y) alkyl” refers to substituted or unsubstitutedsaturated hydrocarbon groups, including straight-chain alkyl andbranched-chain alkyl groups that contain from x to y carbons in thechain, including haloalkyl groups. Preferred haloalkyl groups includetrifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl, andpentafluoroethyl. C₀ alkyl indicates a hydrogen where the group is in aterminal position, a bond if internal. The terms “C_(2-y) alkenyl” and“C_(2-y) alkynyl” refer to substituted or unsubstituted unsaturatedaliphatic groups analogous in length and possible substitution to thealkyls described above, but that contain at least one double or triplebond respectively.

The term “alkylamino”, as used herein, refers to an amino groupsubstituted with at least one alkyl group.

The term “alkylthio”, as used herein, refers to a thiol groupsubstituted with an alkyl group and may be represented by the generalformula alkylS—.

The term “alkynyl”, as used herein, refers to an aliphatic groupcontaining at least one triple bond and is intended to include both“unsubstituted alkynyls” and “substituted alkynyls”, the latter of whichrefers to alkynyl moieties having substituents replacing a hydrogen onone or more carbons of the alkynyl group. Such substituents may occur onone or more carbons that are included or not included in one or moretriple bonds. Moreover, such substituents include all those contemplatedfor alkyl groups, as discussed above, except where stability isprohibitive. For example, substitution of alkynyl groups by one or morealkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups iscontemplated.

The term “amide”, as used herein, refers to a group

wherein each R^(A) independently represent a hydrogen or hydrocarbylgroup, or two R^(A) are taken together with the N atom to which they areattached complete a heterocycle having from 4 to 8 atoms in the ringstructure.

The terms “amine” and “amino” are art-recognized and refer to bothunsubstituted and substituted amines and salts thereof, e.g., a moietythat can be represented by

wherein each R^(A) independently represents a hydrogen or a hydrocarbylgroup, or two R^(A) are taken together with the N atom to which they areattached complete a heterocycle having from 4 to 8 atoms in the ringstructure.

The term “aminoalkyl”, as used herein, refers to an alkyl groupsubstituted with an amino group.

The term “aralkyl”, as used herein, refers to an alkyl group substitutedwith an aryl group.

The term “aryl” as used herein include substituted or unsubstitutedsingle-ring aromatic groups in which each atom of the ring is carbon.Preferably the ring is a 6- or 10-membered ring, more preferably a6-membered ring. The term “aryl” also includes polycyclic ring systemshaving two or more cyclic rings in which two or more carbons are commonto two adjoining rings wherein at least one of the rings is aromatic,e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, aryls,heteroaryls, and/or heterocyclyls. Aryl groups include benzene,naphthalene, phenanthrene, phenol, aniline, and the like.

The term “carbamate” is art-recognized and refers to a group

wherein each R^(A) independently represent hydrogen or a hydrocarbylgroup, such as an alkyl group, or both R^(A) taken together with theintervening atom(s) complete a heterocycle having from 4 to 8 atoms inthe ring structure.

The terms “carbocycle”, and “carbocyclic”, as used herein, refers to asaturated or unsaturated ring in which each atom of the ring is carbon.The term carbocycle includes both aromatic carbocycles and non-aromaticcarbocycles. Non-aromatic carbocycles include both cycloalkane rings, inwhich all carbon atoms are saturated, and cycloalkene rings, whichcontain at least one double bond. “Carbocycle” includes 5-7 memberedmonocyclic and 8-12 membered bicyclic rings. Each ring of a bicycliccarbocycle may be selected from saturated, unsaturated and aromaticrings. Carbocycle includes bicyclic molecules in which one, two or threeor more atoms are shared between the two rings. The term “fusedcarbocycle” refers to a bicyclic carbocycle in which each of the ringsshares two adjacent atoms with the other ring. Each ring of a fusedcarbocycle may be selected from saturated, unsaturated and aromaticrings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, maybe fused to a saturated or unsaturated ring, e.g., cyclohexane,cyclopentane, or cyclohexene. Any combination of saturated, unsaturatedand aromatic bicyclic rings, as valence permits, is included in thedefinition of carbocyclic. Exemplary “carbocycles” include cyclopentane,cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene,1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene andadamantane. Exemplary fused carbocycles include decalin, naphthalene,1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane,4,5,6,7-tetrahydro-1H-indene and bicyclo[4.1.0]hept-3-ene. “Carbocycles”may be susbstituted at any one or more positions capable of bearing ahydrogen atom.

A “cycloalkyl” group is a cyclic hydrocarbon which is completelysaturated. “Cycloalkyl” includes monocyclic and bicyclic rings.Typically, a monocyclic cycloalkyl group has from 3 to about 10 carbonatoms, more typically 3 to 8 carbon atoms unless otherwise defined. Thesecond ring of a bicyclic cycloalkyl may be selected from saturated,unsaturated and aromatic rings. Cycloalkyl includes bicyclic moleculesin which one, two or three or more atoms are shared between the tworings. The term “fused cycloalkyl” refers to a bicyclic cycloalkyl inwhich each of the rings shares two adjacent atoms with the other ring.The second ring of a fused bicyclic cycloalkyl may be selected fromsaturated, unsaturated and aromatic rings. A “cycloalkenyl” group is acyclic hydrocarbon containing one or more double bonds.

The term “carbocyclylalkyl”, as used herein, refers to an alkyl groupsubstituted with a carbocycle group.

The term “carbonate” is art-recognized and refers to a group—OCO₂—R^(A), wherein R^(A) represents a hydrocarbyl group.

The term “carboxy”, as used herein, refers to a group represented by theformula —CO₂H.

The term “ester”, as used herein, refers to a group —C(O)OR whereinR^(A) represents a hydrocarbyl group.

The term “ether”, as used herein, refers to a hydrocarbyl group linkedthrough an oxygen to another hydrocarbyl group. Accordingly, an ethersubstituent of a hydrocarbyl group may be hydrocarbyl-O—. Ethers may beeither symmetrical or unsymmetrical. Examples of ethers include, but arenot limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethersinclude “alkoxyalkyl” groups, which may be represented by the generalformula alkyl-O-alkyl.

The terms “halo” and “halogen” as used herein means halogen and includeschloro, fluoro, bromo, and iodo.

The terms “hetaralkyl” and “heteroaralkyl”, as used herein, refers to analkyl group substituted with a hetaryl group.

The term “heteroalkyl”, as used herein, refers to a saturated orunsaturated chain of carbon atoms and at least one heteroatom, whereinno two heteroatoms are adjacent.

The terms “heteroaryl” and “hetaryl” include substituted orunsubstituted aromatic single ring structures, preferably 5- to7-membered rings, more preferably 5- to 6-membered rings, whose ringstructures include at least one heteroatom, preferably one to fourheteroatoms, more preferably one or two heteroatoms. The terms“heteroaryl” and “hetaryl” also include polycyclic ring systems havingtwo or more cyclic rings in which two or more carbons are common to twoadjoining rings wherein at least one of the rings is heteroaromatic,e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, aryls,heteroaryls, and/or heterocyclyls. Heteroaryl groups include, forexample, pyrrole, furan, thiophene, imidazole, oxazole, thiazole,pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.

The term “heteroatom” as used herein means an atom of any element otherthan carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, andsulfur.

The terms “heterocyclyl”, “heterocycle”, and “heterocyclic” refer tosubstituted or unsubstituted non-aromatic ring structures, preferably 3-to 10-membered rings, more preferably 3- to 7-membered rings, whose ringstructures include at least one heteroatom, preferably one to fourheteroatoms, more preferably one or two heteroatoms. The terms“heterocyclyl” and “heterocyclic” also include polycyclic ring systemshaving two or more cyclic rings in which two or more carbons are commonto two adjoining rings wherein at least one of the rings isheterocyclic, e.g., the other cyclic rings can be cycloalkyls,cycloalkenyls, aryls, heteroaryls, and/or heterocyclyls. Heterocyclylgroups include, for example, piperidine, piperazine, pyrrolidine,tetrahydropyran, tetrahydrofuran, morpholine, lactones, lactams, and thelike.

The term “heterocyclylalkyl” or “heterocycloalkyl”, as used herein,refers to an alkyl group substituted with a heterocycle group.

The term “hydrocarbyl”, as used herein, refers to a group that is bondedthrough a carbon atom that does not have a ═O or ═S substituent, andtypically has at least one carbon-hydrogen bond and a primarily carbonbackbone, but may optionally include heteroatoms. Thus, groups likemethyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered to behydrocarbyl for the purposes of this application, but substituents suchas acetyl (which has a ═O substituent on the linking carbon) and ethoxy(which is linked through oxygen, not carbon) are not. Hydrocarbyl groupsinclude, but are not limited to aryl, heteroaryl, carbocycle,heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

The term “hydroxyalkyl”, as used herein, refers to an alkyl groupsubstituted with a hydroxy group.

The term “lower” when used in conjunction with a chemical moiety, suchas, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant toinclude groups where there are ten or fewer non-hydrogen atoms in thesubstituent, preferably six or fewer. A “lower alkyl”, for example,refers to an alkyl group that contains ten or fewer carbon atoms,preferably six or fewer. In certain embodiments, acyl, acyloxy, alkyl,alkenyl, alkynyl, or alkoxy substituents defined herein are respectivelylower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, orlower alkoxy, whether they appear alone or in combination with othersubstituents, such as in the recitations hydroxyalkyl and aralkyl (inwhich case, for example, the atoms within the aryl group are not countedwhen counting the carbon atoms in the alkyl substituent).

The terms “polycyclyl”, “polycycle”, and “polycyclic” refer to two ormore rings (e.g., cycloalkyls, cycloalkenyls, aryls, heteroaryls, and/orheterocyclyls) in which two or more atoms are common to two adjoiningrings, e.g., the rings are “fused rings”. Each of the rings of thepolycycle can be substituted or unsubstituted. In certain embodiments,each ring of the polycycle contains from 3 to 10 atoms in the ring,preferably from 5 to 7.

The term “silyl” refers to a silicon moiety with three hydrocarbylmoieties attached thereto.

The term “substituted” refers to moieties having substituents replacinga hydrogen on one or more carbons of the backbone. It will be understoodthat “substitution” or “substituted with” includes the implicit provisothat such substitution is in accordance with permitted valence of thesubstituted atom and the substituent, and that the substitution resultsin a stable compound, e.g., which does not spontaneously undergotransformation such as by rearrangement, cyclization, elimination, etc.As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, aromatic and non-aromaticsubstituents of organic compounds. The permissible substituents can beone or more and the same or different for appropriate organic compounds.For purposes of this invention, the heteroatoms such as nitrogen mayhave hydrogen substituents and/or any permissible substituents oforganic compounds described herein which satisfy the valences of theheteroatoms. Substituents can include any substituents described herein,for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, analkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as athioester, a thioacetate, or a thioformate), an alkoxy, a phosphoryl, aphosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine,an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, asulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, aheterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. Inpreferred embodiments, the substituents on substituted alkyls areselected from C₁₋₆ alkyl, C₁₋₆ cycloalkyl, halogen, carbonyl, cyano, orhydroxyl. In more preferred embodiments, the substituents on substitutedalkyls are selected from fluoro, carbonyl, cyano, or hydroxyl. It willbe understood by those skilled in the art that substituents canthemselves be substituted, if appropriate. Unless specifically stated as“unsubstituted,” references to chemical moieties herein are understoodto include substituted variants. For example, reference to an “aryl”group or moiety implicitly includes both substituted and unsubstitutedvariants.

The term “sulfate” is art-recognized and refers to the group —OSO₃H, ora pharmaceutically acceptable salt thereof.

The term “sulfonamide” is art-recognized and refers to the grouprepresented by the general formulae

wherein each R^(A) independently represents hydrogen or hydrocarbyl,such as alkyl, or both R^(A) taken together with the intervening atom(s)complete a heterocycle having from 4 to 8 atoms in the ring structure.

The term “sulfoxide” is art-recognized and refers to the group—S(O)—R^(A), wherein R^(A) represents a hydrocarbyl.

The term “sulfonate” is art-recognized and refers to the group SO₃H, ora pharmaceutically acceptable salt thereof.

The term “sulfone” is art-recognized and refers to the group—S(O)₂—R^(A), wherein R^(A) represents a hydrocarbyl.

The term “thioalkyl”, as used herein, refers to an alkyl groupsubstituted with a thiol group.

The term “thioester”, as used herein, refers to a group —C(O)SR^(A) or—SC(O)R^(A) wherein R^(A) represents a hydrocarbyl.

The term “thioether”, as used herein, is equivalent to an ether, whereinthe oxygen is replaced with a sulfur.

The term “urea” is art-recognized and may be represented by the generalformula

wherein each R^(A) independently represents hydrogen or a hydrocarbyl,such as alkyl, or any occurrence of R^(A) taken together with anotherand the intervening atom(s) complete a heterocycle having from 4 to 8atoms in the ring structure.

“Protecting group” refers to a group of atoms that, when attached to areactive functional group in a molecule, mask, reduce or prevent thereactivity of the functional group. Typically, a protecting group may beselectively removed as desired during the course of a synthesis.Examples of protecting groups can be found in Greene and Wuts,Protective Groups in Organic Chemistry, 3^(rd) Ed., 1999, John Wiley &Sons, NY and Harrison et al., Compendium of Synthetic Organic Methods,Vols. 1-8, 1971-1996, John Wiley & Sons, NY. Representative nitrogenprotecting groups include, but are not limited to, formyl, acetyl,trifluoroacetyl, benzyl, benzyloxycarbonyl (“CBZ”), tert-butoxycarbonyl(“Boc”), trimethylsilyl (“TMS”), 2-trimethylsilyl-ethanesulfonyl(“TES”), trityl and substituted trityl groups, allyloxycarbonyl,9-fluorenylmethyloxycarbonyl (“FMOC”), nitro-veratryloxycarbonyl(“NVOC”) and the like. Representative hydroxyl protecting groupsinclude, but are not limited to, those where the hydroxyl group iseither acylated (esterified) or alkylated such as benzyl and tritylethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilylethers (e.g., TMS or TIPS groups), glycol ethers, such as ethyleneglycol and propylene glycol derivatives and allyl ethers.

As used herein, a therapeutic that “prevents” a disorder or conditionrefers to a compound that, in a statistical sample, reduces theoccurrence of the disorder or condition in the treated sample relativeto an untreated control sample, or delays the onset or reduces theseverity of one or more symptoms of the disorder or condition relativeto the untreated control sample.

The term “treating” includes prophylactic and/or therapeutic treatments.The term “prophylactic or therapeutic” treatment is art-recognized andincludes administration to the host of one or more of the subjectcompositions. If it is administered prior to clinical manifestation ofthe unwanted condition (e.g., disease or other unwanted state of thehost animal) then the treatment is prophylactic (i.e., it protects thehost against developing the unwanted condition), whereas if it isadministered after manifestation of the unwanted condition, thetreatment is therapeutic, (i.e., it is intended to diminish, ameliorate,or stabilize the existing unwanted condition or side effects thereof).

The phrases “conjoint administration” and “administered conjointly”refer to any form of administration of two or more different therapeuticcompounds such that the second compound is administered while thepreviously administered therapeutic compound is still effective in thebody (e.g., the two compounds are simultaneously effective in thepatient, which may include synergistic effects of the two compounds).For example, the different therapeutic compounds can be administeredeither in the same formulation or in a separate formulation, eitherconcomitantly or sequentially. In certain embodiments, the differenttherapeutic compounds can be administered within one hour, 12 hours, 24hours, 36 hours, 48 hours, 72 hours, or a week of one another. Thus, anindividual who receives such treatment can benefit from a combinedeffect of different therapeutic compounds.

The term “prodrug” is intended to encompass compounds which, underphysiologic conditions, are converted into the therapeutically activeagents of the present invention. A common method for making a prodrug isto include one or more selected moieties which are hydrolyzed underphysiologic conditions to reveal the desired molecule. In otherembodiments, the prodrug is converted by an enzymatic activity of thehost animal. For example, esters or carbonates (e.g., esters orcarbonates of alcohols or carboxylic acids) are preferred prodrugs ofthe present invention. In certain embodiments, some or all of thecompounds of the invention in a formulation represented above can bereplaced with the corresponding suitable prodrug, e.g., wherein ahydroxyl in the parent compound is presented as an ester or a carbonateor carboxylic acid present in the parent compound is presented as anester.

As used herein, “small molecules” refers to small organic or inorganicmolecules of molecular weight below about 3,000 Daltons. In general,small molecules useful for the invention have a molecular weight of lessthan 3,000 Daltons (Da). The small molecules can be, e.g., from at leastabout 100 Da to about 3,000 Da (e.g., between about 100 to about 3,000Da, about 100 to about 2500 Da, about 100 to about 2,000 Da, about 100to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 toabout 500 Da, about 200 to about 1500, about 500 to about 1000, about300 to about 1000 Da, or about 100 to about 250 Da).

In some embodiments, a “small molecule” refers to an organic, inorganic,or organometallic compound typically having a molecular weight of lessthan about 1000. In some embodiments, a small molecule is an organiccompound, with a size on the order of 1 nm. In some embodiments, smallmolecule drugs of the invention encompass oligopeptides and otherbiomolecules having a molecular weight of less than about 1000.

An “effective amount” is an amount sufficient to effect beneficial ordesired results. For example, a therapeutic amount is one that achievesthe desired therapeutic effect. This amount can be the same or differentfrom a prophylactically effective amount, which is an amount necessaryto prevent onset of disease or disease symptoms. An effective amount canbe administered in one or more administrations, applications or dosages.A therapeutically effective amount of a composition depends on thecomposition selected. The compositions can be administered from one ormore times per day to one or more times per week; including once everyother day. The skilled artisan will appreciate that certain factors mayinfluence the dosage and timing required to effectively treat a subject,including but not limited to the severity of the disease or disorder,previous treatments, the general health and/or age of the subject, andother diseases present. Moreover, treatment of a subject with atherapeutically effective amount of the compositions described hereincan include a single treatment or a series of treatments.

Compounds of the Invention

One aspect of the invention provides small molecule inhibitors of TRPC5.

In some embodiments, the compound of the invention is a compound ofFormula (I) or (II), or a pharmaceutically acceptable salt thereof;

wherein

-   -   R¹ is selected from the group consisting of alkyl; cycloalkyl;        heterocycloalkyl; aryl; heteroaryl; alkylene-aryl;        alkylene-heteroaryl; alkylene-O-aryl; alkylene-N(alkyl)₂;        alkylene-heterocycloalkyl; alkylene-cycloalkyl; —N(alkyl)₂; and        —C(O)-aryl;    -   R² is selected from the group consisting of alkyl; cycloalkyl;        heterocycloalkyl; aryl; heteroaryl; alkylene-N(alkyl)₂;        alkylene-heterocycloalkyl; alkylene-cycloalkyl;        alkylene-heterocycloalkyl; and alkylene-OR′;    -   R³ is independently selected from alkyl, halogen, OMe, OH,        N(Me)₂, CF₃, OCF₃, CHF₂, OCHF₂, and —O-alkylene-OH;    -   R′ is H, methyl, ethyl, or isopropyl; and    -   n is 0, 1, 2, 3, or 4;    -   provided that the compound is not

In some embodiments, R¹ is selected from the group consisting of methyl,ethyl, n-propyl, i-propyl, n-butyl, i-butyl, and t-butyl. In someembodiments, R¹ is cycloalkyl or heterocycloalkyl.

In some embodiments, R¹ is aryl or heteroaryl.

In some embodiments, R¹ is alkylene-aryl. In some embodiments,alkylene-aryl is methylene-aryl. In some embodiments, methylene-aryl ismethylene-substituted aryl. In some embodiments, methylene-aryl ismethylene-phenyl. In some embodiments, methylene-phenyl ismethylene-substituted phenyl. In some embodiments, substituted phenyl issubstituted with one or more substituent independently selected fromalkyl, halogen, CN, OMe, OH, NO₂, NH₂, N(Me)₂, CF₃, OCF₃, CHF₂, andOCHF₂.

In some embodiments, R¹ is alkylene-N(Me)₂ or alkylene-N(Et)₂. In someembodiments, alkylene-N(Me)₂ is ethylene-N(Me)₂. In some embodiments,alkylene-N(Et)₂ is ethylene-N(Et)₂.

In some embodiments, R¹ is —N(Me)₂ or —N(Et)₂.

In some embodiments, R¹ is alkylene-heteroaryl. In some embodiments,alkylene-heteroaryl is methylene-heteroaryl. In some embodiments,methylene-heteroaryl is methylene-heteroary, wherein heteroaryl is a 5or 6 membered-ring comprising one N atom. In some embodiments,methylene-heteroaryl is methylene-heteroary, wherein heteroaryl is a 5or 6 membered-ring comprising two N atoms. In some embodiments, theheteroaryl is substituted with one or more substituent independentlyselected from alkyl, halogen, CN, OMe, OH, NO₂, NH₂, N(Me)₂, CF₃, OCF₃,CHF₂, and OCHF₂. In some embodiments, methylene-heteroaryl ismethylene-pyridine. In some embodiments, methylene-pyridine ismethylene-substituted pyridine.

In some embodiments, R¹ is alkylene-O-aryl. In some embodiments,alkylene-(O)-aryl is alkylene-(O)-phenyl. In some embodiments,alkylene-(O)-phenyl is alkylene-(O)-substituted phenyl. In someembodiments, alkylene-(O)-phenyl is methylene-(O)-phenyl.

In some embodiments, R¹ is alkylene-heterocycloalkyl. In someembodiments, alkylene-heterocycloalkyl is methylene-heterocycloalkyl. Insome embodiments, heterocycloalkyl is a 3-6 membered-ring.

In some embodiments, R¹ is alkylene-heterocycloalkyl. In someembodiments, alkylene-heterocycloalkyl is alkylene-substitutedheterocycloalkyl. In some embodiments, alkylene heterocylcoalkyl is

wherein Y is N, O, or S; m is an integer selected from 0, 1, 2, or 3,and

represents a 4-12 membered heterocycle. In some embodiments, the 4-12membered heterocycle comprising one or more heteroatoms. In someembodiments,

In some embodiments,

In some embodiments, alkylene heterocylcoalkyl is

wherein Y is N, O, or S; m is an integer selected from 0, 1, 2, or 3,and

represents a 4-12 membered heterocycle. In some embodiments, the 4-12membered heterocycle comprising one or more heteroatoms. In someembodiments,

In some embodiments

In some embodiments, R¹ is alkylene-cycloalkyl. In some embodiments,alkylene-cycloalkyl is methylene-cycloalkyl. In some embodiments,cycloalkyl is a 3-6 membered-ring.

In some embodiments, R¹ is —C(O)-phenyl. In some embodiments, —C(O)-arylis —C(O)— phenyl. In some embodiments, —C(O)-phenyl is —C(O)-substitutedphenyl.

In some embodiments, R² is alkyl. In some embodiments, alkyl is methyl,ethyl, n-propyl, i-propyl, n-butyl, i-butyl, or t-butyl. In someembodiments, R² is t-butyl.

In some embodiments, R² is selected from the group consisting of methyl,ethyl, n-propyl, i-propyl, n-butyl, i-butyl, and t-butyl. In someembodiments, R² is cycloalkyl or heterocycloalkyl.

In some embodiments, R² is aryl or heteroaryl.

In some embodiments, R² is alkylene-N(alkyl)₂. In some embodiments,alkylene-N(alkyl)₂ is alkylene-N(methyl)₂. In some embodiments,alkylene-N(alkyl)₂ is alkylene-N(ethyl)₂. In some embodiments,alkylene-N(alkyl)₂ is methylene-N(alkyl)₂. In some embodiments,alkylene-N(alkyl)₂ is ethylene-N(alkyl)₂.

In some embodiments, R² is alkylene-heterocycloalkyl. In someembodiments, alkylene-heterocycloalkyl is methylene-heterocycloalkyl. Insome embodiments, alkylene-heterocycloalkyl isethylene-heterocycloalkyl. In some embodiments, heterocycloalkyl is a3-6 membered-ring.

In some embodiments, R² is alkylene-heterocycloalkyl. In someembodiments, alkylene heterocylcoalkyl is

wherein Y is N, O, or S; m is an integer selected from 0, 1, 2, or 3,and

represents a 4-12 membered heterocycle. In some embodiments, the 4-12membered heterocycle comprisin one or more heteroatoms. In someembodiments

In some embodiments,

In some embodiments, alkylene heterocylcoalkyl is

wherein Y is N, O, or S; m is an integer selected from 0, 1, 2, or 3,and

represents a 4-12 membered heterocycle. In some embodiments, the 4-12membered heterocycle comprising one or more heteroatoms. In someembodiments,

In some embodiments,

In some embodiments, R² is alkylene-cycloalkyl. In some embodiments,alkylene-cycloalkyl is methylene-cycloalkyl. In some embodiments,alkylene-cycloalkyl is ethylene-cycloalkyl. In some embodiments,cycloalkyl is a 3-6 membered-ring.

In some embodiments, R² is R² is alkylene-OH. In some embodiments, R² isR² is alkylene-OH. In some embodiments, alkylene-OH is methylene-OH. Insome embodiments, -alkylene-OH is ethylene-OH. In some embodiments,alkylene-OH is i-propylene-OH.

In some embodiments, R³ is alkyl. In some embodiments, alkyl is methyl,ethyl, n-propyl, i-propyl, n-butyl, i-butyl, or t-butyl. In someembodiments, R³ is methyl.

In some embodiments, R³ is halogen. In some embodiments, R³ is F. Insome embodiments, R³ is Cl.

In some embodiments, R³ is —O-(alkylene)-OH. In some embodiments,—O-(alkylene)-OH is —O—(CH₂)₂—OH.

In some embodiments, n is 0. In some embodiments, n is 1. In someembodiments, n is 2.

In some embodiments, the compound is selected from the group consistingof:

In some embodiments, the compound is selected from the group consistingof:

In some embodiments, the compound is selected from the group consistingof:

In some embodiments, the compound is selected from the group consistingof:

In some embodiments, the compound is selected from the group consistingof:

In some embodiments, the compound is selected from the group consistingof:

in certain embodiments, the compounds of the invention may be racemic.In certain embodiments, the compounds of the invention may be enrichedin one enantiomer. For example, a compound of the invention may havegreater than 30% ee, 40% ee, 50% ee, 60% ee, 70% ee, 80% ee, 90% ee, oreven 95% or greater ee.

The compounds of the invention have more than one stereocenter.Accordingly, the compounds of the invention may be enriched in one ormore diastereomers. For example, a compound of the invention may havegreater than 30% de, 40% de, 50% de, 60% de, 70% de, 80% de, 90% de, oreven 95% or greater de. In certain embodiments, the compounds of theinvention have substantially one isomeric configuration at one or morestereogenic centers, and have multiple isomeric configurations at theremaining stereogenic centers.

In certain embodiments, the enantiomeric excess of the stereocenter isat least 40% ee, 50% ee, 60% ee, 70% ee, 80% ee, 90% ee, 92% ee, 94% ee,95% ee, 96% ee, 98% ee or greater ee.

As used herein, single bonds drawn without stereochemistry do notindicate the stereochemistry of the compound.

As used herein, hashed or bolded non-wedge bonds indicate relative, butnot absolute, stereochemical configuration (e.g., do not distinguishbetween enantiomers of a given diastereomer).

As used herein, hashed or bolded wedge bonds indicate absolutestereochemical configuration.

In certain embodiments, a therapeutic preparation of the compound of theinvention may be enriched to provide predominantly one enantiomer of acompound. An enantiomerically enriched mixture may comprise, forexample, at least 60 mol percent of one enantiomer, or more preferablyat least 75, 90, 95, or even 99 mol percent. In certain embodiments, thecompound enriched in one enantiomer is substantially free of the otherenantiomer, wherein substantially free means that the substance inquestion makes up less than 10%, or less than 5%, or less than 4%, orless than 3%, or less than 2%, or less than 1% as compared to the amountof the other enantiomer, e.g., in the composition or compound mixture.For example, if a composition or compound mixture contains 98 grams of afirst enantiomer and 2 grams of a second enantiomer, it would be said tocontain 98 mol percent of the first enantiomer and only 2% of the secondenantiomer.

In certain embodiments, a therapeutic preparation may be enriched toprovide predominantly one diastereomer of the compound of the invention.A diastereomerically enriched mixture may comprise, for example, atleast 60 mol percent of one diastereomer, or more preferably at least75, 90, 95, or even 99 mol percent

Methods of Treatment

The non-selective Ca²⁺-permeable Transient Receptor Potential (TRP)channels act as sensors that transduce extracellular cues to theintracellular environment in diverse cellular processes, including actinremodeling and cell migration (Greka et al., Nat Neurosci 6, 837-845,2003; Ramsey et al., Annu Rev Physiol 68, 619-647, 2006; Montell,Pflugers Arch 451, 19-28, 2005; Clapham, Nature 426, 517-524, 2003).Dynamic rearrangement of the actin cytoskeleton relies onspatiotemporally regulated Ca²⁺ influx (Zheng and Poo, Annu Rev Cell DevBiol 23, 375-404, 2007); Brandman and Meyer, Science 322, 390-395,2008); Collins and Meyer, Dev Cell 16, 160-161, 2009) and the smallGTPases RhoA and Rac1 serve as key modulators of these changes(Etienne-Manneville and Hall, Nature 420, 629-635, 2002); Rafiopoulouand Hall, Dev Biol 265, 23-32, 2004). RhoA induces stress fiber andfocal adhesion formation, while Rac1 mediates lamellipodia formation(Etienne-Manneville and Hall, Nature 420, 629-635, 2002). The TransientReceptor Potential Cation Channel, subfamily C, member 5 (TRPC5) acts inconcert with TRPC6 to regulate Ca2+ influx, actin remodeling, and cellmotility in kidney podocytes and fibroblasts. TRPC5-mediated Ca²⁺ influxincreases Rac1 activity, whereas TRPC6-mediated Ca2+ influx promotesRhoA activity. Gene silencing of TRPC6 channels abolishes stress fibersand diminishes focal contacts, rendering a motile, migratory cellphenotype. In contrast, gene silencing of TRPC5 channels rescues stressfiber formation, rendering a contractile cell phenotype. The resultsdescribed herein unveil a conserved signaling mechanism whereby TRPC5and TRPC6 channels control a tightly regulated balance of cytoskeletaldynamics through differential coupling to Rac1 and RhoA.

Ca²⁺-dependent remodeling of the actin cytoskeleton is a dynamic processthat drives cell migration (Wei et al., Nature 457, 901-905, 2009). RhoAand Rac1 act as switches responsible for cytoskeletal rearrangements inmigrating cells (Etienne-Manneville and Hall, Nature 420, 629-635,2002); Raftopoulou and Hall, Dev Biol 265, 23-32, 2004). Activation ofRac1 mediates a motile cell phenotype, whereas RhoA activity promotes acontractile phenotype (Etienne-Manneville and Hall, Nature 420, 629-635,2002). Ca²⁺ plays a central role in small GTPase regulation (Aspenstromet al., Biochem J 377, 327-337, 2004). Spatially and temporallyrestricted flickers of Ca²⁺ are enriched near the leading edge ofmigrating cells (Wei et al., Nature 457, 901-905, 2009).Ca2+microdomains have thus joined local bursts in Rac1 activity(Gardiner et al., Curr Biol 12, 2029-2034, 2002; Machacek et al., Nature461,99-103, 2009) as critical events at the leading edge. To date, thesources of Ca2+influx responsible for GTPase regulation remain largelyelusive. TRP (Transient Receptor Potential) channels generate time andspace-limited Ca²⁺ signals linked to cell migration in fibroblasts andneuronal growth cones0. Specifically, TRPC5 channels are knownregulators of neuronal growth cone guidancel and their activity inneurons is dependent on PI3K and Rac1 activity (Bezzerides et al., NatCell Biol 6, 709-720, 2004).

Podocytes are neuronal-like cells that originate from the metanephricmesenchyme of the kidney glomerulus and are essential to the formationof the kidney filtration apparatus (Somlo and Mundel, Nat Genet. 24,333-335, 2000; Fukasawa et al., J Am Soc Nephrol 20, 1491-1503, 2009).Podocytes possess an exquisitely refined repertoire of cytoskeletaladaptations to environmental cues (Somlo and Mundel, Nat Genet 24,333-335, 2000, Garg et al., Mol Cell Biol 27, 8698-8712, 2007; Verma etal., J Clin Invest 116, 1346-1359, 2006; Verma et al., J Biol Chem 278,20716-20723, 2003; Barletta et al., J Biol Chem 278, 19266-19271, 2003;Holzman et al., Kidney Int 56, 1481-1491, 1999; Ahola et al., Am JPathol 155, 907-913, 1999; Tryggvason and Wartiovaara, N Engl J Med 354,1387-1401, 2006; Schnabel and Farquhar, J Cell Biol 111, 1255-1263,1990; Kurihara et al., Proc Nat Acad Sci USA 89, 7075-7079, 1992). Earlyevents of podocyte injury are characterized by dysregulation of theactin cytoskeleton (Faul et al., Trends Cell Biol 17, 428-437, 2007;Takeda et al., J Clin Invest 108, 289-301, 2001; Asanuma et al., NatCell Biol 8, 485-491, 2006) and Ca2+ homeostasis (Hunt et al., J Am SocNephrol 16,1593-1602, 2005; Faul et al., Nat Med 14, 931-938, 2008).These changes are associated with the onset of proteinuria, the loss ofalbumin into the urinary space, and ultimately kidney failure(Tryggvason and Wartiovaara, N Engl J Med 354, 1387-1401, 2006). Thevasoactive hormone Angiotensin U induces Ca²⁺ influx in podocytes, andprolonged treatment results in loss of stress fibers (Hsu et al., J MolMed 86, 1379-1394, 2008). While there is a recognized link between Ca2+influx and cytoskeletal reorganization, the mechanisms by which thepodocyte senses and transduces extracellular cues that modulate cellshape and motility remain elusive. TRP Canonical 6 (TRPC6) channelmutations have been linked to podocyte injury (Winn et al., Science 308,1801-1804, 2005; Reiser et al., Nat Genet 37, 739-744, 2005; Moller etal., J Am Soc Nephrol 18, 29-36, 2007; Hsu et al., Biochim Biophys Acta1772, 928-936, 2007), but little is known about the specific pathwaysthat regulate this process. Moreover, TRPC6 shares close homology withsix other members of the TRPC channel family (Ramsey et al., Annu RevPhysiol 68, 619-647, 2006; Clapham, Nature 426, 517-524, 2003). TRPC5channels antagonize TRPC6 channel activity to control a tightlyregulated balance of cytoskeletal dynamics through differential couplingto distinct small GTPases.

Proteinuria

Proteinuria is a pathological condition wherein protein is present inthe urine. Albuminuria is a type of proteinuria. Microalbuminuria occurswhen the kidney leaks small amounts of albumin into the urine. In aproperly functioning body, albumin is not normally present in urinebecause it is retained in the bloodstream by the kidneys.Microalbuminuria is diagnosed either from a 24-hour urine collection (20to 200 μg/min) or, more commonly, from elevated concentrations (30 to300 mg/L) on at least two occasions. Microalbuminuria can be aforerunner of diabetic nephropathy. An albumin level above these valuesis called macroalbuminuria. Subjects with certain conditions, e.g.,diabetic nephropathy, can progress from microalbuminuria tomacroalbuminuria and reach a nephrotic range (>3.5 g/24 hours) as kidneydisease reaches advanced stages.

Causes of Proteinuria

Proteinuria can be associated with a number of conditions, includingfocal segmental glomerulosclerosis, IgA nephropathy, diabeticnephropathy, lupus nephritis, membranoproliferative glomerulonephritis,progressive (crescentic) glomerulonephritis, and membranousglomerulonephritis.

A. Focal Segmental Glonmerulosclerosis (FSGS)

Focal Segmental Glomerulosclerosis (FSGS) is a disease that attacks thekidney's filtering system (glomeruli) causing serious scarring. FSGS isone of the many causes of a disease known as Nephrotic Syndrome, whichoccurs when protein in the blood leaks into the urine (proteinuria).

Very few treatments are available for patients with FSGS. Many patientsare treated with steroid regimens, most of which have very harsh sideeffects. Some patients have shown to respond positively toimmunosuppressive drugs as well as blood pressure drugs which have shownto lower the level of protein in the urine. To date, there is nocommonly accepted effective treatment or cure and there are no FDAapproved drugs to treat FSGS. Therefore, more effective methods toreduce or inhibit proteinuria are desirable.

B. IgA Nephropathy

IgA nephropathy (also known as IgA nephritis, IgAN, Berger's disease,and synpharyngitic glomerulonephritis) is a form of glomerulonephritis(inflammation of the glomeruli of the kidney). IgA nephropathy is themost common glomerulonephritis throughout the world. Primary IgAnephropathy is characterized by deposition of the IgA antibody in theglomerulus. There are other diseases associated with glomerular IgAdeposits, the most common being Henoch-Schnlein purpura (HSP), which isconsidered by many to be a systemic form of IgA nephropathy.Henoch-Schönlein purpura presents with a characteristic purpuric skinrash, arthritis, and abdominal pain and occurs more commonly in youngadults (16-35 yrs old). HSP is associated with a more benign prognosisthan IgA nephropathy. In IgA nephropathy there is a slow progression tochronic renal failure in 25-30% of cases during a period of 20 years.

C. Diabetic Nephropathy

Diabetic nephropathy, also known as Kimmelstiel-Wilson syndrome andintercapillary glomerulonephritis, is a progressive kidney diseasecaused by angiopathy of capillaries in the kidney glomeruli. It ischaracterized by nephrotic syndrome and diffuse glomerulosclerosis. Itis due to longstanding diabetes mellitus and is a prime cause fordialysis. The earliest detectable change in the course of diabeticnephropathy is a thickening in the glomerulus. At this stage, the kidneymay start allowing more serum albumin than normal in the urine. Asdiabetic nephropathy progresses, increasing numbers of glomeruli aredestroyed by nodular glomerulosclerosis and the amount of albuminexcreted in the urine increases.

D. Lupus Nephritis

Lupus nephritis is a kidney disorder that is a complication of systemiclupus erythematosus. Lupus nephritis occurs when antibodies andcomplement build up in the kidneys, causing inflammation. It oftencauses proteinuria and may progress rapidly to renal failure. Nitrogenwaste products build up in the bloodstream. Systemic lupus erythematosuscauses various disorders of the internal structures of the kidney,including interstitial nephritis. Lupus nephritis affects approximately3 out of 10,000 people.

E. Membranoproliferative Glomerulonephritis I/II/III

Membranoproliferative glomerulonephritis is a type of glomerulonephritiscaused by deposits in the kidney glomerular mesangium and basementmembrane thickening, activating complement and damaging the glomeruli.There are three types of membranoproliferative glomerulonephritis. TypeI is caused by immune complexes depositing in the kidney and is believedto be associated with the classical complement pathway. Type II issimilar to Type I, however, it is believed to be associated with thealternative complement pathway. Type III is very rare and it ischaracterized by a mixture of subepithelial deposits and the typicalpathological findings of Type I disease.

F. Progressive (Crescentic) Glomerulonephritis

Progressive (crescentic) glomerulonephritis (PG) is a syndrome of thekidney that, if left untreated, rapidly progresses into acute renalfailure and death within months. In 50% of cases, PG is associated withan underlying disease such as Goodpasture's syndrome, systemic lupuserythematosus, or Wegener granulomatosis; the remaining cases areidiopathic. Regardless of the underlying cause, PG involves severeinjury to the kidney's glomeruli, with many of the glomeruli containingcharacteristic crescent-shaped scars. Patients with PG have hematuria,proteinuria, and occasionally, hypertension and edema. The clinicalpicture is consistent with nephritic syndrome, although the degree ofproteinuria may occasionally exceed 3 g/24 hours, a range associatedwith nephrotic syndrome. Untreated disease may progress to decreasedurinary volume (oliguria), which is associated with poor kidneyfunction.

G. Membranous Glomerulonephritis

Membranous glomerulonephritis (MGN) is a slowly progressive disease ofthe kidney affecting mostly patients between ages of 30 and 50 years,usually Caucasian. It can develop into nephrotic syndrome. MGN is causedby circulating immune complex. Current research indicates that themajority of the immune complexes are formed via binding of antibodies toantigens in situ to the glomerular basement membrane. The said antigensmay be endogenous to the basement membrane, or deposited from systemiccirculation.

Measurement of Urine Protein Levels

Protein levels in urine can be measured using methods known in the art.Until recently, an accurate protein measurement required a 24-hour urinecollection. In a 24-hour collection, the patient urinates into acontainer, which is kept refrigerated between trips to the bathroom. Thepatient is instructed to begin collecting urine after the first trip tothe bathroom in the morning. Every drop of urine for the rest of the dayis to be collected in the container. The next morning, the patient addsthe first urination after waking and the collection is complete.

More recently, researchers have found that a single urine sample canprovide the needed information. In the newer technique, the amount ofalbumin in the urine sample is compared with the amount of creatinine, awaste product of normal muscle breakdown. The measurement is called aurine albumin-to-creatinine ratio (UACR). A urine sample containing morethan 30 milligrams of albumin for each gram of creatinine (30 mg/g) is awarning that there may be a problem. If the laboratory test exceeds 30mg/g, another UACR test should be performed 1 to 2 weeks later. If thesecond test also shows high levels of protein, the person has persistentproteinuria, a sign of declining kidney function, and should haveadditional tests to evaluate kidney function.

Tests that measure the amount of creatinine in the blood will also showwhether a subject's kidneys are removing wastes efficiently. Too muchcreatinine in the blood is a sign that a person has kidney damage. Aphysician can use the creatinine measurement to estimate how efficientlythe kidneys are filtering the blood. This calculation is called theestimated glomerular filtration rate, or eGFR. Chronic kidney disease ispresent when the eGFR is less than 60 milliliters per minute (mL/min).

TRPC5

TRPC is a family of transient receptor potential cation channels inanimals. TRPC5 is subtype of the TRPC family of mammalian transientreceptor potential ion channels. Three examples of TRPC5 are highlightedbelow in Table 1.

TABLE 1 The TRPC5 orthologs from three different species along withtheir GenBank Ref Seq Accession Numbers. Species Nucleic Acid Amino AcidGeneID Homo sapiens NM_012471.2 NP_036603.1 7224 Mus musculusNM_009428.2 NP_033454.1 22067 Rattus norvegicus NM_080898.2 NP_543174.1140933

Accordingly, in certain embodiments, the invention provides methods fortreating, or the reducing risk of developing, a kidney diseasecomprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of the invention (e.g., a compound ofFormula I), or a pharmaceutical composition comprising said compound.

In some embodiments, the kidney disease is selected from the groupconsisting of Focal Segmental Glomerulosclerosis (FSGS), Diabeticnephropathy, Alport syndrome, hypertensive kidney disease, nephroticsyndrome, steroid-resistant nephrotic syndrome, minimal change disease,membranous nephropathy, idiopathic membranous nephropathy,membranoproliferative glomerulonephritis (MPGN), immune complex-mediatedMPGN, complement-mediated MPGN, Lupus nephritis, postinfectiousglomerulonephritis, thin basement membrane disease, mesangialproliferative glomerulonephritis, amyloidosis (primary), c1qnephropathy, rapidly progressive GN, anti-GBM disease, C3glomerulonephritis, hypertensive nephrosclerosis, and IgA nephropathy.In some embodiments, the kidney disease is proteinuria. In someembodiments, the kidney disease is microalbuminuria or macroalbuminuria.

The invention also provides methods of treating, or the reducing risk ofdeveloping, anxiety, or depression, or cancer, comprising administeringto a subject in need thereof a therapeutically effective amount of acompound of the invention (e.g., a compound of Formula I), or apharmaceutical composition comprising said compound.

Subjects to be Treated

In one aspect of the invention, a subject is selected on the basis thatthey have, or are at risk of developing, a kidney disease, anxiety,depression, or cancer.

Subjects that have, or are at risk of developing, proteinuria includethose with diabetes, hypertension, or certain family backgrounds. In theUnited States, diabetes is the leading cause of end-stage renal disease(ESRD). In both type 1 and type 2 diabetes, albumin in the urine is oneof the first signs of deteriorating kidney function. As kidney functiondeclines, the amount of albumin in the urine increases. Another riskfactor for developing proteinuria is hypertension. Proteinuria in aperson with high blood pressure is an indicator of declining kidneyfunction. If the hypertension is not controlled, the person can progressto full kidney failure. African Americans are more likely thanCaucasians to have high blood pressure and to develop kidney problemsfrom it, even when their blood pressure is only mildly elevated. Othergroups at risk for proteinuria are American Indians, Hispanics/Latinos,Pacific Islander Americans, older adults, and overweight subjects.

In one aspect of the invention, a subject is selected on the basis thatthey have, or are at risk of developing proteinuria. A subject that has,or is at risk of developing, proteinuria is one having one or moresymptoms of the condition. Symptoms of proteinuria are known to those ofskill in the art and include, without limitation, large amounts ofprotein in the urine, which may cause it to look foamy in the toilet.Loss of large amounts of protein may result in edema, where swelling inthe hands, feet, abdomen, or face may occur. These are signs of largeprotein loss and indicate that kidney disease has progressed. Laboratorytesting is the only way to find out whether protein is in a subject'surine before extensive kidney damage occurs.

The methods are effective for a variety of subjects including mammals,e.g., humans and other animals, such as laboratory animals, e.g., mice,rats, rabbits, or monkeys, or domesticated and farm animals, e.g., cats,dogs, goats, sheep, pigs, cows, or horses. In some embodiments, thesubject is a mammal. In some embodiments, the subject is a human.

EXAMPLES

The invention is further described in the following examples, which donot limit the scope of the invention described in the claims.

Example 1: Synthetic Methods

The following illustrate synthetic routes to exemplary compounds of theinvention.

Example 2: TRPCx Assay Protocols

I. Fluorescence-Based Assays

A. TRPC4

HEK 293 cells expressing human TRPC4 cells were trypsinised, counted andseeded in black, clear-bottomed 96-well plates at a density of 50,000cells per well and incubated overnight Next day, the cells were loadedwith membrane potential dye. Dye solution was made up according to themanufacturer's instructions in HEPES buffered Hank's balanced saltsolution (HBSS). Dye solution (10 μL) was added to the wells andincubated at 37f for 1 hour. The test compounds and standard inhibitorswere added to the wells and incubated at room temperature for 10minutes. The plates were then placed in the flexstation and fluorescencemonitored every 1.52 seconds. After 20 seconds, 10 μL of the appropriatestandard agonist was added and the fluorescence monitored for 2 minutesat ex/emm: 530 nm/565 nm

B. TRPC5

HEK 293 cells expressing human TRPC5 cells were trypsinised, counted andseeded in black, clear-bottomed 96-well plates at a density of 50,000cells per well and incubated overnight Next day, the cells were loadedwith membrane potential dye. Dye solution was made up according to themanufacturer's instructions in HEPES buffered Hank's balanced saltsolution (HBSS). Dye solution (10 μL) was added to the wells andincubated at 37° C. for 1 hour. The test compounds and standardinhibitors were added to the wells and incubated at room temperature for10 minutes. The plates were then placed in the flexstation andfluorescence monitored every 1.52 seconds. After 20 seconds, 10 μL ofthe appropriate standard agonist was added and the fluorescencemonitored for 2 minutes at ex/emm: 530 nm/565 nm.

C. TRPC6

HEK 293 cells expressing human TRPC6 cells were trypsinised, counted andseeded in black, clear-bottomed 96-well plates at a density of 50,000cells per well and incubated overnight Next day, the cells were loadedwith membrane potential dye (Molecular Devices, cat: R8127). Dyesolution was made up according to the manufacturer's instructions inHEPES buffered Hank's balanced salt solution (HBSS). Dye solution wasadded to the wells and incubated at 37° C. for 1 hour. The testcompounds and standard inhibitors were added to the wells and incubatedat room temperature for 10 minutes prior to addition of activator. Theplates were then placed in the flexstation and fluorescence monitoredevery 1.52 seconds. After 20 seconds, the standard activator (Carbachol)was added and the fluorescence monitored for 2 minutes at ex/emm: 530nm/565 nm.

II. Automated Patch Clamp Assay (Qpatch)

A. TRPC5

HEK 293 cells expressing human TRPC5 were harvested, re-suspended inserum free medium, and added to the automated platform and used within2-3 hours. Internal and external physiological solutions were freshlyprepared prior to the assay. The external solution contained: 145 mMNaCl, 4 mM KCl, 2 mM CaCl₂), 1 mM MgCl2, 10 mM HEPES, 10 mM glucose, pH7.4 with NaOH and 300 mOsm/L. The internal solution contained 120 mML-aspartic acid, 120 mM CsOH.H2O, 20 mM CsCl, 2 mM MgCl2, 8.8 mM CaCl₂),10 mM EGTA, 10 mM HEPES, 10 mM Glucose, 0.1 mM GTP and 2 mM Na2ATP; pH7.2 with CsOH and 290 mOsm/L. The free internal Ca2+ concentration wasbuffered to 1 μM.

The automated electrophysiological platform QPatch 16 from Sophion(Denmark) was used to carry out the compounds profiling. The seriesresistance and quality of seals were continuously monitored during theexperiments. Data was analyzed using Sophion QPatch assay software 5.6(Odense). IC50 values were calculated using a least squares regressionalgorithm (Hill equation).

To monitor the ion currents, a voltage ramp from −100 mV to +100 mV,over 300 ms, was applied every 10 seconds, from a holding potential of−60 mV.

After recording for a minimum of 60 seconds control period,Rosiglitazone (30 uM), was applied to activate the channel.

B. TRPC4

HEK 293 cells expressing human TRPC4 were harvested, re-suspended inserum free medium, added to the automated platform and used within 2-3hours. Internal and external physiological solutions were freshlyprepared prior the assay. The external solution contained: 145 mM NaCl,4 mM KCl, 2 mM CaCl2), 1 mM MgCl2, 10 mM HEPES, 10 mM glucose, pH 7.4with NaOH and 300 mOsm/L. The internal solution contained 120 mML-aspartic acid, 120 mM CsOH.H2O, 20 mM CsCl, 2 mM MgCl2, 10 mM EGTA, 10mM HEPES, 10 mM Glucose, and 2 mM Na2ATP; pH 7.2 with CsOH and 290mOsm/L.

The TRPC4 channel agonist Englerin was used to activate and assess testcompounds.

The automated electrophysiological platform Qpatch 16 from Sophion(Denmark) was used to carry out the compounds profiling. The seriesresistance and quality of seals were continuously monitored during theexperiments. Data was analyzed using Sophion Qpatch assay software 5.6(Odense) and Microsoft Office Excel 2007. IC50 values were calculatedusing a least squares regression algorithm (Hill equation).

C. TRPC6

HEK 293 cells expressing human TRPC6 were harvested, re-suspended inserum free medium, added to the automated platform and used within 2-3hours. Internal and external physiological solutions were freshlyprepared prior the assay. The external solution contained: 145 mM NaCl,4 mM KCl, 2 mM CaCl2), 1 mM MgCl2, 10 mM HEPES, 10 mM glucose, pH 7.4with NaOH and 300 mOsm/L. The internal solution contained 120 mML-aspartic acid, 120 mM CsOH.H2O, 20 mM CsCl, 2 mM MgCl2, 10 mM EGTA, 10mM HEPES, 10 mM Glucose, and 2 mM Na2ATP; pH 7.2 with CsOH and 290mOsm/L.

The agonist OAG EC50 was used to activate TRPC6 and assess testcompounds.

The automated electrophysiological platform Qpatch 16 from Sophion(Denmark) was used to carry out the compounds profiling. The seriesresistance and quality of seals were continuously monitored during theexperiments. Data was analyzed using Sophion Qpatch assay software 5.6(Odense) and Microsoft Office Excel 2007. IC50 values were calculatedusing a least squares regression algorithm (Hill equation).

Example 3: Exemplary Biological Assay Data

TABLE 2 IC₅₀ values for representative 1-substituted 7-azaindoles of thedisclosure measured in an automated patch clamp assay utilizing HEK293cells overexpressing TRPC5 (see above), with the readout as a currentblock utilizing whole cell automated patch following stimulation withrosiglitazone at either 80 or 100 mV. TRPC5 IC₅₀ TRPC5 IC₅₀ Structure 80mV (μM) 100 mV (μM)

3.51

6.24 4.19

7.07

13.85 8.52

16.95 16.57

18.3 21.62

19.6 19.12

23.62 25.17

>30

>30

>30 >30

>30 >30

>30 >30

>30

>30 >30

>30

>30 >30

>30

>30 >30

>30 >30

>30

>30 >30

>30

>30 >30

>30 >30

>30 >30

>30 >30

>30 >30

>30

>30 >30

>30 >30

>30

>30

>30

>30

>30

>30

>30 >30

>30

>30 >30

>30

>30 >30

>30 >30

>30

>30

>30 >30

>30 >30

>30 >30

>30

>30

>30 >30

>30

>30

>30

>30

>30

>30

>30

>30

>30

TABLE 3 IC₅₀ values for representative 7-substituted 7-azaindoles of thedisclosure measured in an automated patch clamp assay utilizing HEK293cells overexpressing TRPC5 (see above), with the readout as a currentblock utilizing whole cell automated patch following stimulation withrosiglitazone at 100 mV. TRPC5 IC₅₀ Structures 100 mV (μM)

5.47

6.02

7.64

8.1

8.57

9.79

10.18

10.38

12.2

14

14.38

15.65

17.65

22.23

24.51

>30

What is claimed is:
 1. A compound of Formula (I) or (II), or apharmaceutically acceptable salt thereof;

wherein R¹ is selected from the group consisting of alkyl; cycloalkyl;heterocycloalkyl; aryl; heteroaryl; alkylene-aryl; alkylene-heteroaryl;alkylene-O-aryl; alkylene-N(alkyl)₂; alkylene-heterocycloalkyl;alkylene-cycloalkyl; —N(alkyl)₂, and —C(O)-aryl; R² is selected from thegroup consisting of alkyl; cycloalkyl; heterocycloalkyl; aryl;heteroaryl; alkylene-N(alkyl)₂; alkylene-heterocycloalkyl;alkylene-cycloalkyl; alkylene-heterocycloalkyl; and alkylene-OR′; R³ isindependently selected from alkyl, halogen, OMe, OH, N(Me)₂, CF₃, OCF₃,CHF₂, OCHF₂, and —O-alkylene-OH; R′ is H, methyl, ethyl, or isopropyl;and n is 0, 1, 2, 3, or 4; provided that the compound is not


2. The compound of claim 1, wherein R′ is methylene-aryl.
 3. Thecompound of claim 2, wherein aryl is phenyl.
 4. The compound of claim 2,wherein aryl is substituted phenyl.
 5. The compound of claim 4, whereinthe phenyl is substituted with one or more substituent independentlyselected from alkyl, halogen, CN, OMe, OH, NO₂, NH₂, N(Me)₂, CF₃, OCF₃,CHF₂, and OCHF₂.
 6. The compound of claim 1, wherein R¹ ismethylene-heteroaryl.
 7. The compound of claim 6, wherein heteroaryl isa 5 or 6 membered-ring comprising one N atom.
 8. The compound of claim6, wherein heteroaryl is a 5 or 6 membered-ring comprising two N atoms.9. The compound of any one of claims 6-8, wherein the heteroaryl issubstituted with one or more substituent independently selected fromalkyl, halogen, CN, OMe, OH, NO₂, NH₂, N(Me)₂, CF₃, OCF₃, CHF₂, andOCHF₂.
 10. The compound of claim 1, wherein R¹ is alkylene-O-aryl. 11.The compound of claim 10, wherein alkylene-O-aryl is methylene-O-phenyl.12. The compound of claim 1, wherein R¹ is methylene-heterocycloalkyl.13. The compound of claim 1, wherein heterocycloalkyl is a 3-6membered-ring.
 14. The compound of claim 1, wherein R¹ ismethylene-cycloalkyl.
 15. The compound of claim 1, wherein cycloalkyl isa 3-6 membered-ring.
 16. The compound of claim 1, wherein R¹ is—C(O)-phenyl.
 17. The compound of any one of claims 1-16, wherein R² isselected from the group consisting of methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, and t-butyl.
 18. The compound of claim 17, wherein R²is t-butyl.
 19. The compound of any one of claims 1-16, wherein R² isalkylene-N(methyl)₂ or alkylene-N(ethylene)₂.
 20. The compound of claim19, wherein alkylene is methylene or ethylene.
 21. The compound of anyone of claims 1-16, wherein R² is selected fromalkylene-heterocycloalkyl; alkylene-cycloalkyl; andalkylene-heterocycloalkyl.
 22. The compound of claim 21, whereinalkylene is methylene or ethylene.
 23. The compound of any one of claims1-16, wherein R² is alkylene-OH.
 24. The compound of any one of claims1-23, wherein R³ is methyl.
 25. The compound of any one of claims 1-23,wherein R³ is F or Cl.
 26. The compound of any one of claims 1-23,wherein R³ is —O—(CH₂)₂—OH.
 27. The compound of any one of claims 1-23,wherein n is
 0. 28. The compound of any one of claims 1-26, wherein nis
 1. 29. The compound of any one of claims 1-26, wherein n is
 2. 30.The compound of claim 1, wherein the compound is selected from the groupconsisting of:


31. The compound of claim 1, wherein the compound is selected from thegroup consisting of:


32. The compound of claim 1, wherein the compound is selected from thegroup consisting of:


33. The compound of claim 1, wherein the compound is selected from thegroup consisting of:


34. The compound of claim 1, wherein the compound is selected from thegroup consisting of:


35. The compound of claim 1, wherein the compound is selected from thegroup consisting of:


36. A composition, comprising a compound of any one of claims 1-35 or apharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable excipient.
 37. A method of treating, or the reducing risk ofdeveloping, a kidney disease, anxiety, or depression, or cancer,comprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of any one of claims 1-35.
 38. The methodof claim 37, wherein a kidney disease is treated or the risk ofdeveloping a kidney disease is reduced.
 39. The method of claim 37,wherein a kidney disease is treated.
 40. The method of any one of claims37-39, wherein the kidney disease is selected from the group consistingof Focal Segmental Glomerulosclerosis (FSGS), Diabetic nephropathy,Alport syndrome, hypertensive kidney disease, nephrotic syndrome,steroid-resistant nephrotic syndrome, minimal change disease, membranousnephropathy, idiopathic membranous nephropathy, membranoproliferativeglomerulonephritis (MPGN), immune complex-mediated MPGN,complement-mediated MPGN, Lupus nephritis, postinfectiousglomerulonephritis, thin basement membrane disease, mesangialproliferative glomerulonephritis, amyloidosis (primary), c1qnephropathy, rapidly progressive GN, anti-GBM disease, C3glomerulonephritis, hypertensive nephrosclerosis, and IgA nephropathy.41. The method of any one of claims 37-39, wherein the kidney disease isproteinuria.
 42. The method of any one of claims 37-39, wherein thekidney disease is microalbuminuria or macroalbuminuria.
 43. The methodof any one of claims 37-42, wherein the subject is a mammal
 44. Themethod of claim 43, wherein the mammal is a human.